Deficient repair response of IPF fibroblasts in a co-culture model of epithelial injury and repair

Fibrogenesis & Tissue Repair

Gabor Jarai²  Cory M Hogaboam¹  Sony Prasad² 

[1] Department of Medicine, Cedars Sinai Medical Center, AHSP Room A9108, 127 S. San Vicente Blvd, Los Angeles, CA 90048-3311, USA;Novartis Institutes of Biomedical Research, Respiratory Disease Area, Wimblehurst Road, Horsham, West Sussex RH12 5AB, UK

关键词: PDGF;    HGF;    bFGF;    PDGFR;    Wound healing;    Co-culture;    IPF;    Idiopathic pulmonary fibrosis;   

Others  :  802268
DOI  :  10.1186/1755-1536-7-7

 received in 2014-01-16, accepted in 2014-04-17,  发布年份 2014

【 摘 要 】

Background

Idiopathic pulmonary fibrosis (IPF) is a progressive disorder marked by relentless fibrosis and damage of the lung architecture. A growing body of evidence now suggests that IPF progresses as a result of aberrant epithelial-fibroblast crosstalk. Injured epithelia are a major source of growth factors such as PDGF which guide resident fibroblasts to injury sites.

Results

In this study, we utilized a novel co-culture system to investigate the effect of fibroblast phenotype on their response to epithelial injury. Fibroblasts from normal lungs (NHLF) responded to epithelial injury and populated the wound site forming a fibroblast plug/mechanical barrier which prevented epithelial wound closure. IPF fibroblasts were impaired in their response to epithelial injury. They also expressed reduced PDGFRα compared to NHLFs and were defective towards PDGF-AA mediated directional movement. Neutralization of PDGF-AA and pan-PDGF but not PDGF-BB reduced the injury response of NHLFs thereby preventing the formation of the mechanical barrier and promoting epithelial wound closure. Co-culture of epithelial cells with IPF fibroblasts led to marked increase in the levels of pro-fibrotic growth factors - bFGF and PDGF and significant depletion of anti-fibrotic HGF in the culture medium. Furthermore, IPF fibroblasts but not NHLFs induced a transient increase in mesenchymal marker expression in the wound lining epithelial cells. This was accompanied by increased migration and faster wound closure in co-cultures with IPF fibroblasts.

Conclusions

Our data demonstrate that the IPF fibroblasts have an aberrant repair response to epithelial injury.

【 授权许可】

   
2014 Prasad et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Farkas L, Gauldie J, Voelkel NF, Kolb M: Pulmonary hypertension and idiopathic pulmonary fibrosis: a tale of angiogenesis, apoptosis, and growth factors. Am J Respir Cell Mol Biol 2011, 45:1-15.
• [2]Eickelberg O, Laurent GJ: The quest for the initial lesion in idiopathic pulmonary fibrosis: gene expression differences in IPF fibroblasts. Am J Respir Cell Mol Biol 2010, 42:1-2.
• [3]Qunn L, Takemura T, Ikushima S, Ando T, Yanagawa T, Akiyama O, Oritsu M, Tanaka N, Kuroki T: Hyperplastic epithelial foci in honeycomb lesions in idiopathic pulmonary fibrosis. Virchows Arch 2002, 441:271-278.
• [4]Allen JT, Spiteri MA: Growth factors in idiopathic pulmonary fibrosis: relative roles. Respir Res 2002, 3:13. BioMed Central Full Text
• [5]Hinz B, Phan SH, Thannickal VJ, Prunotto M, Desmouliere A, Varga J, De WO, Mareel M, Gabbiani G: Recent developments in myofibroblast biology: paradigms for connective tissue remodeling. Am J Pathol 2012, 180:1340-1355.
• [6]Phan SH: Genesis of the myofibroblast in lung injury and fibrosis. Proc Am Thorac Soc 2012, 9:148-152.
• [7]Moodley YP, Caterina P, Scaffidi AK, Misso NL, Papadimitriou JM, McAnulty RJ, Laurent GJ, Thompson PJ, Knight DA: Comparison of the morphological and biochemical changes in normal human lung fibroblasts and fibroblasts derived from lungs of patients with idiopathic pulmonary fibrosis during FasL-induced apoptosis. J Pathol 2004, 202:486-495.
• [8]Gardner A, Borthwick LA, Fisher AJ: Lung epithelial wound healing in health and disease. Expert Rev Respir Med 2010, 4:647-660.
• [9]Ramos C, Montano M, Garcia-Alvarez J, Ruiz V, Uhal BD, Selman M, Pardo A: Fibroblasts from idiopathic pulmonary fibrosis and normal lungs differ in growth rate, apoptosis, and tissue inhibitor of metalloproteinases expression. Am J Respir Cell Mol Biol 2001, 24:591-598.
• [10]Marchand-Adam S, Marchal J, Cohen M, Soler P, Gerard B, Castier Y, Leseche G, Valeyre D, Mal H, Aubier M, Dehoux M, Crestani B: Defect of hepatocyte growth factor secretion by fibroblasts in idiopathic pulmonary fibrosis. Am J Respir Crit Care Med 2013, 168:1156-1161.
• [11]Standiford TJ, Rolfe MR, Kunkel SL, Lynch JP III, Becker FS, Orringer MB, Phan S, Strieter RM: Altered production and regulation of monocyte chemoattractant protein-1 from pulmonary fibroblasts isolated from patients with idiopathic pulmonary fibrosis. Chest 1993, 103:121S.
• [12]Cruz-Gervis R, Stecenko AA, Dworski R, Lane KB, Loyd JE, Pierson R, King G, Brigham KL: Altered prostanoid production by fibroblasts cultured from the lungs of human subjects with idiopathic pulmonary fibrosis. Respir Res 2002, 3:17. BioMed Central Full Text
• [13]Moodley YP, Scaffidi AK, Misso NL, Keerthisingam C, McAnulty RJ, Laurent GJ, Mutsaers SE, Thompson PJ, Knight DA: Fibroblasts isolated from normal lungs and those with idiopathic pulmonary fibrosis differ in interleukin-6/gp130-mediated cell signaling and proliferation. Am J Pathol 2003, 163:345-354.
• [14]Selman M, Pardo A: Idiopathic pulmonary fibrosis: misunderstandings between epithelial cells and fibroblasts? Sarcoidosis Vasc Diffuse Lung Dis 2004, 21:165-172.
• [15]Rogel MR, Soni PN, Troken JR, Sitikov A, Trejo HE, Ridge KM: Vimentin is sufficient and required for wound repair and remodeling in alveolar epithelial cells. FASEB J 2011, 25:3873-3883.
• [16]Walter MN, Wright KT, Fuller HR, MacNeil S, Johnson WE: Mesenchymal stem cell-conditioned medium accelerates skin wound healing: an in vitro study of fibroblast and keratinocyte scratch assays. Exp Cell Res 2010, 316:1271-1281.
• [17]Schmidt BA, Horsley V: Intradermal adipocytes mediate fibroblast recruitment during skin wound healing. Development 2013, 140:1517-1527.
• [18]Kanazawa S, Fujiwara T, Matsuzaki S, Shingaki K, Taniguchi M, Miyata S, Tohyama M, Sakai Y, Yano K, Hosokawa K, Kubo T: bFGF regulates PI3-kinase-Rac1-JNK pathway and promotes fibroblast migration in wound healing. PLoS One 2010, 5:e12228.
• [19]Schneider L, Cammer M, Lehman J, Nielsen SK, Guerra CF, Veland IR, Stock C, Hoffmann EK, Yoder BK, Schwab A, Satir P, Christensen ST: Directional cell migration and chemotaxis in wound healing response to PDGF-AA are coordinated by the primary cilium in fibroblasts. Cell Physiol Biochem 2010, 25:279-292.
• [20]Bowden DH, Young L, Adamson IY: Fibroblast inhibition does not promote normal lung repair after hyperoxia. Exp Lung Res 1994, 20:251-262.
• [21]Adamson IY, Hedgecock C, Bowden DH: Epithelial cell-fibroblast interactions in lung injury and repair. Am J Pathol 1990, 137:385-392.
• [22]Adamson IY, Young L, Bowden DH: Relationship of alveolar epithelial injury and repair to the induction of pulmonary fibrosis. Am J Pathol 1988, 130:377-383.
• [23]Inoue Y, King TE Jr, Tinkle SS, Dockstader K, Newman LS: Human mast cell basic fibroblast growth factor in pulmonary fibrotic disorders. Am J Pathol 1996, 149:2037-2054.
• [24]Khalil N, Xu YD, O’Connor R, Duronio V: Proliferation of pulmonary interstitial fibroblasts is mediated by transforming growth factor-beta1-induced release of extracellular fibroblast growth factor-2 and phosphorylation of p38 MAPK and JNK. J Biol Chem 2005, 280:43000-43009.
• [25]Crestani B, Marchand-Adam S, Quesnel C, Plantier L, Borensztajn K, Marchal J, Mailleux A, Soler P, Dehoux M: Hepatocyte growth factor and lung fibrosis. Proc Am Thorac Soc 2012, 9:158-163.
• [26]Gregory PA, Bracken CP, Smith E, Bert AG, Wright JA, Roslan S, Morris M, Wyatt L, Farshid G, Lim YY, Lindeman GJ, Shannon MF, Drew PA, Khew-Goodall Y, Goodall GJ: An autocrine TGF-beta/ZEB/miR-200 signaling network regulates establishment and maintenance of epithelial-mesenchymal transition. Mol Biol Cell 2011, 22:1686-1698.
• [27]Homma S, Nagaoka I, Abe H, Takahashi K, Seyama K, Nukiwa T, Kira S: Localization of platelet-derived growth factor and insulin-like growth factor I in the fibrotic lung. Am J Respir Crit Care Med 1995, 152:2084-2089.
• [28]Martin P: Wound healing–aiming for perfect skin regeneration. Science 1997, 276:75-81.
• [29]Martin P, Hopkinson-Woolley J, McCluskey J: Growth factors and cutaneous wound repair. Prog Growth Factor Res 1992, 4:25-44.
• [30]Bonner JC: Regulation of PDGF and its receptors in fibrotic diseases. Cytokine Growth Factor Rev 2004, 15:255-273.
• [31]Agren MS, Eaglstein WH, Ferguson MW, Harding KG, Moore K, Saarialho-Kere UK, Schultz GS: Causes and effects of the chronic inflammation in venous leg ulcers. Acta Derm Venereol Suppl (Stockh) 2000, 210:3-17.
• [32]Mendez MV, Stanley A, Park HY, Shon K, Phillips T, Menzoian JO: Fibroblasts cultured from venous ulcers display cellular characteristics of senescence. J Vasc Surg 1998, 28:876-883.
• [33]Trengove NJ, Bielefeldt-Ohmann H, Stacey MC: Mitogenic activity and cytokine levels in non-healing and healing chronic leg ulcers. Wound Repair Regen 2000, 8:13-25.
• [34]Cisneros J, Hagood J, Checa M, Ortiz-Quintero B, Negreros M, Herrera I, Ramos C, Pardo A, Selman M: Hypermethylation-mediated silencing of p14(ARF) in fibroblasts from idiopathic pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol 2012, 303:L295-L303.
• [35]Coward WR, Watts K, Feghali-Bostwick CA, Knox A, Pang L: Defective histone acetylation is responsible for the diminished expression of cyclooxygenase 2 in idiopathic pulmonary fibrosis. Mol Cell Biol 2009, 29:4325-4339.
• [36]Coward WR, Watts K, Feghali-Bostwick CA, Jenkins G, Pang L: Repression of IP-10 by interactions between histone deacetylation and hypermethylation in idiopathic pulmonary fibrosis. Mol Cell Biol 2010, 30:2874-2886.
• [37]Sanders YY, Pardo A, Selman M, Nuovo GJ, Tollefsbol TO, Siegal GP, Hagood JS: Thy-1 promoter hypermethylation: a novel epigenetic pathogenic mechanism in pulmonary fibrosis. Am J Respir Cell Mol Biol 2008, 39:610-618.
• [38]Sanders YY, Tollefsbol TO, Varisco BM, Hagood JS: Epigenetic regulation of thy-1 by histone deacetylase inhibitor in rat lung fibroblasts. Am J Respir Cell Mol Biol 2011, 45:16-23.
• [39]Antoniades HN, Galanopoulos T, Neville-Golden J, O’Hara CJ: Malignant epithelial cells in primary human lung carcinomas coexpress in vivo platelet-derived growth factor (PDGF) and PDGF receptor mRNAs and their protein products. Proc Natl Acad Sci U S A 1992, 89:3942-3946.
• [40]Kolar M, Szabo P, Dvorankova B, Lacina L, Gabius HJ, Strnad H, Sachova J, Vlcek C, Plzak J, Chovanec M, Cada Z, Betka J, Fik Z, Paces J, Kovarova H, Motlik J, Jarkovska K, Smetana K Jr: Upregulation of IL-6, IL-8 and CXCL-1 production in dermal fibroblasts by normal/malignant epithelial cells in vitro: Immunohistochemical and transcriptomic analyses. Biol Cell 2012, 104:738-751.
• [41]Lee JG, Kay EP: FGF-2-mediated signal transduction during endothelial mesenchymal transformation in corneal endothelial cells. Exp Eye Res 2006, 83:1309-1316.
• [42]Lee HT, Lee JG, Na M, Kay EP: FGF-2 induced by interleukin-1 beta through the action of phosphatidylinositol 3-kinase mediates endothelial mesenchymal transformation in corneal endothelial cells. J Biol Chem 2004, 279:32325-32332.
• [43]Fischer AN, Fuchs E, Mikula M, Huber H, Beug H, Mikulits W: PDGF essentially links TGF-beta signaling to nuclear beta-catenin accumulation in hepatocellular carcinoma progression. Oncogene 2007, 26:3395-3405.
• [44]Stoll SW, Rittie L, Johnson JL, Elder JT: Heparin-binding EGF-like growth factor promotes epithelial-mesenchymal transition in human keratinocytes. J Invest Dermatol 2012, 132:2148-2157.
• [45]Borok Z: Role for alpha3 integrin in EMT and pulmonary fibrosis. J Clin Invest 2009, 119:7-10.
• [46]Kasai H, Allen JT, Mason RM, Kamimura T, Zhang Z: TGF-beta1 induces human alveolar epithelial to mesenchymal cell transition (EMT). Respir Res 2005, 6:56. BioMed Central Full Text
• [47]Park J, Kim DS, Shim TS, Lim CM, Koh Y, Lee SD, Kim WS, Kim WD, Lee JS, Song KS: Lung cancer in patients with idiopathic pulmonary fibrosis. Eur Respir J 2001, 17:1216-1219.
• [48]Jakubzick C, Choi ES, Carpenter KJ, Kunkel SL, Evanoff H, Martinez FJ, Flaherty KR, Toews GB, Colby TV, Travis WD, Joshi BH, Puri RK, Hogaboam CM: Human pulmonary fibroblasts exhibit altered interleukin-4 and interleukin-13 receptor subunit expression in idiopathic interstitial pneumonia. Am J Pathol 2004, 164:1989-2001.